Designing of RNA Molecule Translating for Activitable Melit-tin as Selective Targeting of Leishmania Infected Cells
Background: Leishmaniasis is characterized by strong inflammatory responses with high levels of inflammatory cytokines that induce microRNA 21 and matrix metalloproteinases. Melittin has inhibitory effects on proliferation of various cells via induction of apoptosis. Melittin can be integrated in cell membranes and induce apoptosis. Thus, designation of biomolecules for the selective destroy of the infected cells is a treatment option. One approach is the precise engineering of constructs for the selective expression of melittin in the infected cells.
Methods: For this aim we designed a construct composing melittin nucleotide sequence and nucleotide sequence coding for polyanionic peptide function inhibitory element to further guarantee the selective function of melittin in inflamed tissues and infected cells, were included in a construct as melittin inhibitor via matrix metalloproteinase degradable linker.
Results: Reverse complementary sequences were designed so melittin sequences for the selective targeting of Leishmania could be expressed in infected cells using cell microRNA machinery.
Conclusion: Translation machinery in infected cells with increased miR-21 could translate melittin, MMP linker and polyanionic inhibitor through a non-canonical pathway. Then, the MMP linker is degraded and selective killing of Leishmania infected cells would happen.
2. Eiras DP, Kirkman LA, Murray HW. Cu-taneous leishmaniasis: current treatment practices in the USA for returning travel-ers. Curr Treat Options Infect Dis. 2015; 7(1): 52–62.
3. Van Den Bogaart G, Guzman JV, Mika JT, Poolman B. On the mechanism of pore formation by melittin. J Biol Chem. 2008; 283(49): 33854–7.
4. Zarrinnahad H, Mahmoodzadeh A, Ha-midi MP, Mahdavi M, Moradi A, Bagheri KP, Shahbazzadeh D. Apoptotic effect of melittin purified from Iranian honey bee venom on human cervical cancer hela cell line. International journal of peptide re-search and therapeutics. 2018 Dec 1;24(4):563-70.
5. Mahmoodzadeh A, Zarrinnahad H, Bagheri KP, Moradia A, Shahbazzadeh D. First report on the isolation of melittin from Iranian honey bee venom and evalu-ation of its toxicity on gastric cancer AGS cells. J Chin Med Assoc. 2015; 78(10): 574–83.
6. Imai S, Kumar P, Hellen CU, D'Souza VM, Wagner G. An accurately preor-ganized IRES RNA structure enables eIF4G capture for initiation of viral trans-lation. Nat Struct Mol Biol. 2016; 23(9): 859-64.
7. Lewin B, Dover G. Genes v. Oxford: Ox-ford University Press; 1994.
8. Chen SJ, Dill KA. RNA folding energy landscapes. Proc Natl Acad Sci U S A. 2000; 97(2): 646–51.
9. Tinoco Jr I. Force as a useful variable in reactions: unfolding RNA. Annu Rev Bio-phys Biomol Struct. 2004; 33: 363–85.
10. Shin C, Nam JW, Farh KK, Chiang HR, Shkumatava A, Bartel DP. Expanding the microRNA targeting code: functional sites with centered pairing. Mol Cell. 2010; 38(6): 789–802.
11. Wolter JM, Le HH, Linse A, Godlove VA, Nguyen TD, Kotagama K, Lynch A, Rawls A, Mangone M. Evolutionary pat-terns of metazoan microRNAs reveal tar-geting principles in the let-7 and miR-10 families. Genome Res. 2017; 27(1): 53–63.
12. Zhang Z, Hu F, Sung MW, Shu C, Cas-tillo-González C, Koiwa H, Tang G, Dickman M, Li P, Zhang X. RISC-interacting clearing 3’-5’exoribonucleases (RICEs) degrade uridylated cleavage frag-ments to maintain functional RISC in Ara-bidopsis thaliana. Elife. 2017; 6: e24466.
13. Elliott D, Ladomery M. Molecular Biology of RNA (1st ed). Oxford University Press. 2011; pp. 34–64.
14. Meister G, Landthaler M, Patkaniowska A, Dorsett Y, Teng G, Tuschl T. Human Ar-gonaute2 mediates RNA cleavage targeted by miRNAs and siRNAs. Mol Cell. 2004; 15(2): 185–97.
15. Campos TM, Passos ST, Novais FO, Beit-ing DP, Costa RS, Queiroz A, Mosser D, Scott P, Carvalho EM, Carvalho LP. Ma-trix metalloproteinase 9 production by monocytes is enhanced by TNF and par-ticipates in the pathology of human cuta-neous leishmaniasis. Plos Negl Trop Dis. 2014; 8(11): e3282.
16. Cawood R, Chen HH, Carroll F, Bazan-Peregrino M, van Rooijen N, Seymour LW. Use of tissue-specific microRNA to control pathology of wild-type adenovirus without attenuation of its ability to kill can-cer cells. PLoS Pathog. 2009; 5(5): e1000440.
17. Kertesz M, Iovino N, Unnerstall U, Gaul U, Segal E. The role of site accessibility in microRNA target recognition. Nat Genet. 2007; 39(10): 1278.
18. Olson ES, Aguilera TA, Jiang T, Ellies LG, Nguyen QT, Wong EH, Gross LA, Tsien RY. In vivo characterization of activatable cell penetrating peptides for targeting pro-tease activity in cancer. Integr Biol(Camb). 2009; 1(5–6): 382–93.
19. Gondi CS, Lakka SS, Dinh DH, Olivero WC, Gujrati M, Rao JS. Intraperitoneal in-jection of a hairpin RNA–expressing plasmid targeting urokinase-type plasmin-ogen activator (uPA) receptor and uPA re-tards angiogenesis and inhibits intracranial tumor growth in nude mice. Clin Cancer Res. 2007; 13(14): 4051–60.
20. Samadikhah HR, Majidi A, Nikkhah M, Hosseinkhani S. Preparation, characteriza-tion, and efficient transfection of cationic liposomes and nanomagnetic cationic lip-osomes. Int J Nanomedicine. 2011; 6: 2275-2283.
21. Lamiable A, Thevenet P, Rey J, Vavrusa M, Derreumaux P, Tuffery P. PEP-FOLD3: faster de novo structure prediction for linear peptides in solution and in complex. Nu-cleic Acids Res. 2016; 44(W1): W449–W54.
22. Nicholson AW. Ribonuclease III mecha-nisms of double‐stranded RNA cleavage. Wiley Interdiscip Rev RNA. 2014; 5(1):31–48.
23. Woodson SA. Compact intermediates in RNA folding. Annu Rev Biophys. 2010; 39: 61–77.
24. Weenink T, McKiernan RM, Ellis T. ra-tional design of RNA structures that pre-dictably tune eukaryotic gene expression. BioRxiv. 2017; 1: 137877.
25. Androsavich JR, Chau BN, Bhat B, Linsley PS, Walter NG. Disease-linked mi-croRNA-21 exhibits drastically reduced mRNA binding and silencing activity in healthy mouse liver. RNA. 2012; 18(8): 1510–26.
26. Łabno A, Tomecki R, Dziembowski A. Cytoplasmic RNA decay pathways, en-zymes and mechanisms. Biochim Biophys Acta. 2016; 1863(12): 3125–47.
27. Kushner SR. mRNA decay in prokaryotes and eukaryotes: different approaches to a similar problem. IUBMB Life. 2004; 56(10): 585–94.
28. Kandasamy AD, Chow AK, Ali MA, Schulz R. Matrix metalloproteinase-2 and myocardial oxidative stress injury: beyond the matrix. Cardiovasc Res. 2010; 85(3): 413–23.
|Issue||Vol 16 No 3 (2021)|
|RNA design Leishmania spp.; Melittin; miR-21; microRNA machinery|
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